Entamoeba histolytica. Hugo Varet, Yana Shaulov, ... of E. histolytica to NS. Trophozoites (1x106) were treated with (i) GSNO (350 uM) for 1 hour at 37°C (NI);.
SUPPLEMENTARY INFORMATION (9 pages including this) From
Enteric bacteria boost defences against oxidative stress in Entamoeba histolytica Hugo Varet, Yana Shaulov, Odile Sismeiro, Meirav Trebicz-Geffen, Rachel Legendre, Jean-Yves Coppée, Serge Ankri and Nancy Guillen Scientific Reports, 2018 _______________________________________________________________________________________________ Included material Figure S1: Effect of Escherichia coli O55 on the resistance of E. histolytica to nitrosative stress. Figure S2: Effect of diverse bacteria on the resistance of E. histolytica to oxidative stress. Figure S3: GO Terms enrichment in E. histolytica under stress upon incubation with diverse bacteria. Document DOC1: LRR motifs in the amoebic proteins identified in this work (see Table S8).
Figure S1. Effect of Escherichia coli O55 on the resistance of E. histolytica to NS. Trophozoites (1×106) were treated with (i) GSNO (350 μM) for 1 hour at 37°C (NI); (ii) 1×109 live E. coli O55 (LB) for 30 minutes and then with GSNO (350 μM) for 1 hour; and (iii) heat-killed E. coli O55 (KB, the equivalent of 1×109 bacteria) for 30 minutes and then with GSNO (350 μM) for 1 hour. The trophozoites’ viability was determined in an eosin dye exclusion assay.
Figure S2. Effect of diverse bacteria on the resistance of E. histolytica to oxidative stress
120! 100!
Viability (%) !
80! 60! 40! 20! 0!
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Trophozoites (1×10 ) in the presence of the diverse live bacteria (1×10 ) were treated with 2.5 mM H2O2 for 1 hour at 37°C. The trophozoites’ viability was determined in an eosin dye exclusion assay.
Figure S3. GO Terms enrichment in E. histolytica under stress upon incubation with diverse bacteria.
The data from Supplementary Table 6 were analysed with PANTHER tools (http://pantherdb.org/), in order to determine the fold enrichment (FE) of gene sets with GO terms, relative to the background list for all genes in the amoeba genome. The GO terms were used to identify enriched molecular processes; those with an FE ≥ 2.5 were plotted.
Supplementary DOC 1. LRR motifs in the amoebic proteins identified in this work (see Table S8). The diverse LRR motifs were localized by visual inspection of amino acid sequences. We took into account the consensus sequences for LRR Bsp A: LxxIxLxxxVxxIgxxAFxxCxx LRR11: LxxLxLxxNxL LRR12: LxxLxLxxCxxL EHI_120570, 356 aa MQLGYNEIMIVSKYFDDINDFINLEIGVKRFQGNMERFHFNPIPLNQYSRKLFPNIETF HIYNIYDEVFEDGRIFKYVIWYKVDYSRYLEEKEEMNEYKNIEYTQEDRKKYGNTIPNE VTSLGYWCFSLCDDIQEIEIPTSVTEIRSYCFERCSS LQTITIPTNVSKIGDSCFYKCSS LQTITIPTNVSKIRECCFYGCSS LTTINIPPSVTKIGNRCFEECYS LTSINIED-VKYISEKRIFMNEPV LISIKILENLTIINGKNIEKKDINEFIIPSSITKLGDYCFEYCYS LQTINIPTSVSKIGDYCFKECTS LTSINIPSSITSFGKGCFYHCRCEEELKKNKTIPEYCFEE EHI_139980, 666 aa MQRDNPNNMKRIRRKMMSLNIPTRPFLLTENTGSIIPTLSEIILDSFGRFEQNFSSQQE QTPIALPPVLASKLIYRISNNPNASESDLNIILFRILSSGTPLQVIDLHHTHLLTKFTC MNLSQLCNNVSITTLNISYATGISSESLKVLFRSFKQLKYLNTEACVGFDDNAFISLLT IHPPLEVIVISNCPKISDKSVIELKNINSLISFKANNIQLTISSISVLHGLKEIELMGN NCLDDQCLLKISQLNPDLTRICFGNSRLSDEALQRLLQQLNGNLISLNLSGCTRAGPMT LAQLFNSQFKLKYLNLANCFGINGDSILESYHPMFRSTLFRWLEEIEYLNISGCIRLSE EFITTTLIASPQLRSLILDDTAVTDQCLQEFIQSSITYNQTVRRSIHTPGMKVHPLTLS LKRCIRLTDNAYTNLFQTRGVDLISLNLSFCNSLSSLSLNVFSLYSNTLTSFYASNNDM ISEYSWLKFIASCPQLRVLFLSNNRGITNEVINQIKISCPLISHLDISSCLSINQFVIP ILCKMQSLEYIDISFDVMINEDGIISLVNSLTRLSTFCMQGIAFKQSYLFTEEAQWLNS LKLNFIPMCGDDVMRNIASYCPLLTSVELRMCSGVTDNGIEMLLQKCTKISHLVLGGTS ISQFKMMELVGRGLFIA EHI_087890, 484 aa MSSDKQHYNKKNMGIKQFPKDCISQASIITSIDLSNNEIINLPKEMGPFKVLTHFRMMA NKVSSLPLSFTTLTNLRHLDLNANCFTEFPTQITSLTNLEEIQMIQNQLTSIPDCIGNL VKLQRISFTANFLKSLPKGLAKCVDMNYIELTSNEFEEFPDVICELRKVTILMLQQNRI KEVPDSISKLEKLSGLYLSSNNFGKFPESVCTIPSLTQLELDNNNFVDIPDSLSQLTKL KTLIINKSFISCLNSVDMMSNLCQIVLSDTKCMFLPDLSQNSKLTSLNVIRGYLNEVKS LPPNCSCRFSNNQIESIELPENGVLQYMILSNNRLKVSPNLSMLSKISRLDISQNRITR FNENTCHPTLQQLDISCNPLVEFPVCITKCQSLKILNLSDCHLYDIPSNVLSSLSNLET LYIGCNHLSSLESLSVLKKLRALYLQSNNLLHFPQSIFDLITLKTLFVSNNYITTIPNE ISQLTQLEQLAP
